CPC C07K 1/22 (2013.01) [B01D 15/3809 (2013.01); B01D 61/027 (2013.01); B01D 61/145 (2013.01); B01D 61/58 (2013.01); C07K 14/5443 (2013.01); C07K 14/7155 (2013.01); C07K 14/745 (2013.01); C07K 16/36 (2013.01); B01D 2315/16 (2013.01); C07K 2317/565 (2013.01); C07K 2319/00 (2013.01)] | 30 Claims |
1. A method of purifying a multi-chain chimeric polypeptide, the method comprising:
loading an affinity chromatography resin with a liquid comprising the multi-chain chimeric polypeptide;
washing the affinity chromatography resin using one or more wash buffer(s); and
eluting the multi-chain chimeric polypeptide using an elution buffer, wherein:
the affinity chromatography resin comprises an anti-tissue factor antibody or antigen-binding fragment thereof comprising a heavy chain variable domain comprising a CDR1 of SEQ ID NO: 1, a CDR2 of SEQ ID NO: 2 or 9, and a CDR3 of SEQ ID NO: 3, and a light chain variable domain comprising a CDR1 of SEQ ID NO: 4, a CDR2 of SEQ ID NO: 5, and a CDR3 of SEQ ID NO: 6, attached to a base resin;
the multi-chain chimeric polypeptide comprises:
(a) a first chimeric polypeptide comprising:
(i) a first target-binding domain comprising a sequence that is at least 80% identical to amino acids 19 to 305 of SEQ ID NO: 70;
(ii) a soluble tissue factor domain comprising a sequence that is at least 80% identical to SEQ ID NO: 10; and
(iii) a first domain of a pair of affinity domains comprising a sequence that is at least 80% identical to SEQ ID NO: 125; and
(b) a second chimeric polypeptide comprising:
(i) a second domain of a pair of affinity domains comprising a sequence that is at least 80% identical to SEQ ID NO: 123; and
(ii) a second target-binding domain comprising a sequence that is at least 80% identical to amino acids 19 to 305 of SEQ ID NO: 70;
the first chimeric polypeptide and the second chimeric polypeptide associate through the binding of the first domain and the second domain of the pair of affinity domains; and
the multi-chain chimeric polypeptide does not stimulate blood coagulation in a mammal.
|